US3164618A - Alkylated steroids - Google Patents

Description

- 3,164,618 ALKYLATED STERDIDS a Richard Rausser, Union, and Eugene P. Oliveto' field, N..l., assiguors to Schering Corporation, Bloomfield, N.J.', a corporation'of New llersey e No Drawing.

23 vClaims. (Cl. 260-397 .45)

This invention relates to a group of novelialkylated steroid compounds and to methods of preparing the same.

Islama- Filed May s, 1958, Ser; No. 733,843

Patented Jan. 5, 1965 inflammatory properties thereof; Unfortunately, the addition of these 9a-halogens has hitherto resulted in a marked increase in the salt-retaining properties of the known anti-inflammatory steroids, thus preventing their oral or injection administration in many instances, "and making their use by topical application extremely hazardous. We have found unexpectedly, however, thatthe addition of a lo-lower alkyl group, and particularly themethyl radical,

More particularly, this invention relates to 16-alkyl- A -pregnadienes and their halogenated derivatives, as well as certain novel A -pregnenes which are useful in the preparation thereof. t

This application is a continuation-in-part of now aban-. doned US. application, Serial No. 673,141, filed July 23, 1957, by Richard Rausser and Eugene. P. Oliveto, the

I inventors herein, and entitled Alkylated Steroids.

While inhibiting the increasein salt retention effected by the introduction of the 9a-halogen, does not affect the enhanced therapeutic properties of these halo derivatives. Thus, these compounds can now be. administered safely. Similarly, the salt-retaining properties of the non-halogenated steroids, such as, for example,'cortisone and hydrocortisone, are markedly reduced. This is particularly surprising when it is noted that the substitution of a methyl group in the 2- or 6'-position of such known Heretofore it has beenknown to prepare pregnehes such as cortisone and hydrocortisone, and pregnadienes, as, for example, prednisone and prednisolone as well as the corresponding 2l-esters and 9-halo derivatives thereof.

We have now discovered, however, that 16-alkylation of steroids possessing anti-inflammatory properties confers on the parent steroids the pronounced property of nature sis and diuresis. As a result of this unique and unex-' pected property, steroid compounds which po'ssess.,antiinflammatory properties but whichfare limited in their usefulness in the treatment of disease because of their inherent sodium retaining effects have their usefulness extended by16-alkylation, V

The novel compounds of our invention are represented graphically as follows:

. CH3 C In C H2 0 R omenv :b

wherein R is hydrogen or a carboxylic acyl radical and on X R' w steroids; as "9a-fluoro hydrocortisone or 9u-fluoro-prednisolone fails to alleviate their salt retaining characteristic,

which prevents their employment for therapeutic applications such as disclosed herein.

It should be noted that, while the lower alkyl substituent can be introduced into the 16-position by a number of procedures which results in the formation of 160a and 16,8 corticoids and before or after the introduction of the 9a-halogen group, the following represents preferred sequences for preparing the compounds of. our invention. Alternative procedures for introducing the 16-a1ky1 substituent such as, for example, the procedure which comprises reacting a 20-keto-16 dehydro steroid with a nitroalkane, followed by reduction, quaternization and pyrolysis, are also des'cribed hereinafter. Sequences A, B,

preferably one containingfrom 1 to 8 carbon atoms such 1 as, for example, a loweri alkanoyl radical, e.g., acetat propionate, cyclopentyl, 'benzoate, phthalate, dimethyl acetate, 'trimethyl acetate, tert.-butyl acetate, phenoxy acetate, thiophene carboxylate',-nicotinate and like substitu'ents, R is a lower alkyl group such as, for example,

a-isopropyl, B-isopropyl, a-butyl, fi-butyl, a-tertiary (tert.)- butyl, and fi-tert-butyl; X is O, (I-LqOH) and (l-LflOI-I), and Y is hydrogen or a halogen having an atomic weight of less than 81, e.g. bromine, chlorine and fluorine and when X is (H,aOH), Y must be hydrogen. Those compounds wherein -X represents (HaOI-I), although not therapeutically active per se, are convertible by known oxidative procedures into ll-keto compounds which possess physiological activity. Further, as noted above, the dienes as represented by Formula B above, manifest a markedly greater therapeutic activity than the A monoenes of Formula'A. The A -monoenes are, however, also valuable as intermediates in the preparation of the dienes of Formula B above. It is noted that the substitution of an a-h'alogen atom for hydrogen at position .9 in the 16-alkylated glucocorticoids of applicants herein,

will, as with those steroids previously known (e.g. pred- 16-pregnene-3a-ol- IGB-methylpregnanenisone, prednisolone and the like), enhance the antiand C illustrate desirable methods of preparation of the 16,6-alkyl corticoids of our invention; sequence D a practicable procedure for obtaining the Ida-alkyl corticoids of applicants invention; and sequence E a more convenient mode of effecting the production of the 9a-halogen derivatives of the 16a.-alkyl and lfifi-alkyl steroids described in the initial sequences A, B, C and D. For purposes of illustration, the preferred methyl and acetate groups are employed in these sequences, as well as in the subsequent description thereof, as the l'6-alkyl radical, and 2l-acyl substit'uent, respectively, and the 9a-bromo and 9 x-fiuoro substituents as the 9a-halogen radicals.

SEQUENCE A I C Ha 11,20- di0ne 3-acetate SEQUENCE B IGB-methylpregnane- 2,4-dibrom016fi-methyl- -y 17,21-dial-3,11,20-trione pregnaue.17a,21-diol- 21-acetate 3,11,20-tri0ne 21-acetate lfifl-methylprednisone 21-acetate H lfia-rnethylprednisolone acetate,

. 3' SEQUENCE C i I lfia-mcthylprednisone 17a,21-(lihydroxy-16a-mcthyl- 2l-acetate (1) pregnane 3,11,20-trione lfia-methylhydrocortison e lfiasmethyli prednisolone sE UENcE'E- prednisolone 2l-acetate V By way of further illustration, the 1'6,8-lower alkyl (e.g, methyl) compounds of our invention described in reaction hydrocortisone sequence A above are desirably prepared with the easily available substance 16-pregnene-3ix-ol-11,20-dione 3-acylate (e.g'. acetate). Reactioncf this ester with diazomethanegives inexcellent yield, the pyrazoline intermediate, which upon pyrolysis at, or above, its melting point is converted into 16-methyl-16-pregnene-3u-ol-11,20-dione 3-acetate. Where the lee-ethyl, 16,8-propyl, or 165-butyl derivative is desired, for example, the corresponding diazoethane, diazopropan'e, diazobutane or the like is used to other acetylating agents, such diate enol-acetate.

also'bc used in place of acetic anhydride. Furthermore,

the strong acid catalyst is notnecessarily limited to pperoxy-acid, as exemplified by peracetic acid, pe'rbenzoic acid, monoperphthalic acid, pertrifiuoroacetic acid and the. like, Y r

7 Introduction of the 21 acetoxy grou p or other ester thereof is effected in the conventionalmanner, such as by "bromination of the'C-Zl methyl group, followed by reactionof the bromo-Compoundwith, for example, sodium or potassium acetate, whereby 16fl-methylpregnane-3a,17a- ZI-triol-lLZO-dione ZI-acetate is. formed. Although acetoxylation has been shown by Way of illustration, it

will be apparent to one skilled in the art that other acyloxylations,'such as the introduction of a propionoxy group, for example, can be analogously effected. I

in order to obtain the 3-keto-A or 3-keto-A -system,

the saturated pregnane ester l6li-methylpre'gnane-3u,17cc,

Zl-tridl-ILZil-dione Zl-acetate is further-transformed as follows: the hydroxyl group at (1-3 is converted to a keto group, preferably by means of N-bro moacetamide whereby,le-methylpregnane-flai1-diol-3,11,20-trione 21-acetate is produced. It isapparent that'other equivalent oxidizing agents can be employed in this step, such as N- bromosuccinimide, chromium dioxide-pyridine, chromium-trioxide-acetonersulfuric acid mixture {and the like.

. The 'A -double bond is now convenientlyintroduced into the'A-ring by brominating 1fifi-rriethylpregnane-17ot,2l

if 5 'diol-3,11,20-trione- ZI-acetate in the conventional manner 7 whereby the intermediary bromo-compoundis formed,

which may or may not be isolated. Although we prefer to halogenate with bromine, other halogenating-agents such as chlorine are equally effective. Dehydrogenatien of this 4- bromo intermediate, also, in a conventional manner such as by reaction with semicarbazide followed by hydrolysis of the resultant .S-monosemicarbazone, or by refluxing with dimethylformamide optionally in the presence of lithium chloride, or by refluxing with'organic bases suchfas-collidine, causes elimination of hydrogen halide and insertion of the 'A -double bond,yielding liafl-methylcortisone ZI-acetate; This compound can then be saponified to produce the diol-trione, 16B-methylcortisorie, by reacting the former, that is, the l6/3-methyl cortisone 2l-acetate, with hydrolytic agents such as aqueous 1methanolic obtain the corresponding compounds.'--'Although we pre-L fer to pyrolize the pyrazoline by heating above its melting point, such pyrolysis'may also be effected by heating the substance in'a high boiling inert solvent, such as p-cymene, or tctralin and the like.' The unsaturation in the D-ring is conveniently removed by reductive hydrogenation preferably in the presence ofa catalyst, such aspalladium,

whereupon there is obtained l6f3-methylpregnane-3a-ol 11,20-dione 3-acetate. I

In order to introduce a hydroxyl group at C-17,- we

' have found that the 20 keto group of 16,,8-methylpregnane-3a-ol-.1l,20-dione 3-acetate' can be easily converted into an enol-acetate by refluxing with acetic anhydride 1 and a strong acid, such as p-toluenesulfonic acid, for exhydrolyzes the epoxide in situ and thereby forms 16;?

methylpregnane-fm,17a-diol-l1,20-dione, In this procedurc, whereby the hydroxyl group is introduced at C17,

potassium bicarbonate or with such reagents as sodium carbonate, sodium hydroxide, sodium alkoxides (e.g. sodrum ethoxide), and acids such as p-toluenesulfonic acid.

. l'fi-methylcortisone can now be re-esterified at C-21 by introducing any desired acid residue. We have found that, as is generally known in the art, esterification further enhances the duration of activity of the steroid alcohol and provides for acompound which is more eifectively administered parenterally. We have found that acid residues, such as obtained from'acetic acid, propionic acid, trimethylacetic acid, t-butylacetic acid, cyclopentylpropionic acid, furoic acid, phenoxyacetic acid and :the like, provide useful esters. Similarly, half-esters of dibasic acids, such as obtained from phthalic, succinic, tartaric, citric and the-like,provide for esters which can be solubilized by formation of a salt of thefree carboxyl group with an alkali metal, such as sodium. In place of the dibasic organic acids, th ere can be used inorganic acids, such as phosphate and the like, thus producing a dihydrogen phosphate ester which itself can be further solubilized by salt formation.

Introductionof the A -bond is preferably carried out by subjecting methylcortisone Zl-acetate or 165-methylcortisone to the microbiological action of a dehydrog'enating microorganism. We'prefer to utilize Bacillus "sphaericus .(American Type Culture Collection 7055) or Corynebacterium simplex, A.T.C.C .,,l6 '46 [awarding to as isopropenyl acetate, may.

l1,20-trione 2l-acetate.

.bond is inserted can also be accomplished byother known methods, such as by reacting the saturated pregnane ester,

lofi-methylcortisone ZI-acetate, or the pregnane, 16,8- methylpregnane-17u,21-diol-3,l1,20-trione ZI-acetate with i selenium dioxide for example, or with chloranil, at elevated temperatures or by the well-known methodsfof halogenation and dehydrohalogenation. i t

This latter dehydrogenative procedure is exemplified by reaction sequence B whereby the pregnane, l6li-methylpregnane-l7a,2l-diol-3,11,20-trione 21-acetate, is dihalogenated preferably with bromine, thus forming the intermediary; 2,4 dibromo-l6fl-methylpregnane-17a,21-diol-3,

basic agents, preferably dimethylforamide, produces the ZI-acetate of 16,6-methylprednisone which can be saponified by use of any of the hydrolytic agents described above, e.g. potassium bicarbonate, to form the corresponding C-Zl alcohol.- It is obviousjthat at the final stateof l-6B-methylprednisone, various ester groups may be intro- The foregoing reaction sequences and discussions have applied to the preparation of compounds containing a keto function at C-ll. It is apparent that if any of the starting materials contains a hydroxyl group at C-lla, said group will be carried through the series of reactions.

original starting material, 'l6 pregnene-3a l-ll,20-dione 3 -acetate, we prefer to initially carry out the reactions so as to produce ll-keto substances and when required,

reduce the ll-keto group to an ll-hydroxyl group prefi erably having the 5 configuration. Such a transformation Other methods of protecting the 3-keto groups may be employed, such as forming a 3,20-bis-ethylene ketal and then reducing the ll-keto group by means of sodium borohydride, lithium borohydride, or lithium aluminum hydride and the like. Acid hydrolysis of the ll-hydroxybis-ketal so formed yields 16/3-methylhydrocortisone.

16B-methylhydrocortisone can be converted to 16;}

methylprednisolone by means ofthe microbiological fermentation or chemicalmethods described above for the conversion of lofi-methylcortisone to 16,3-methylpredni- However, keeping in mind the ready availability of the l sone. Similarly, 16fl-methylprednisolone can be esterified according to any of the well-known techniques so as to produce the corresponding ZI-acetate, e.g., 16B-methylprednisolone 21*acetate.

Alternatively, 16fi-methylprednisone ZI-acetate itself can be converted to 16B-methylprednisolone 2l-acetate by first selectively protecting the 3,20-diketo groups as described above for the mono-enes and then selectively reducing the ll-keto function.

The 16e-lower alkyl compounds of our invention are desirably prepared by the procedure illustrated in Sequence D above, wherein 3a-hydroxy-l6-pregnene-11,20- dione is added to a standard Grignard reagent, such as I for example, methyl magnesium iodide, prepared from methyl iodide and magnesium, and refluxed to effect the 20 Didhydrobromination with production of the corresponding saturated Ida-methyl derivative, 3u-hydroxy-l6u-methyl pregnanel-' 11,20-dione;

The methyl iodide derivative is againemployedmerely butyl magnesium iodide or the bromide and chlorine.

equivalents thereof ispmployed as the Grignard reagent.

The produce of this Grignard reaction, 3a-hydroxy-l6umethyl pregnane-ll,20-dione, is then hydroxylated' by standard proceduresat the C-17 position, as for example,

by reaction with acetic anhydride and p-toluene sulfonic acid, followed by peracetic acid and then by alkaline hydrolysis to form 3a,17a-dihydroxy 16cc methylpregnane-l 1,20-dione. Introduction of the C-Zl acetoxy group, or other ester thereof, is accomplished by known methods, such as for example,;by bromination of the C-21 methyl group in an inert solvent such as chloroform with the sequential reaction of this product with sodium or potassium acetate in acetone or dimethylformamide. The product thus formed is 3a,17a,21-trihydroxy-16a-methylr pregnane-ll,20-,dione 21'-acetate. The hydroxyl group at position 3 inthis latter compound is then transformed to a keto group,'preferably by means of N-bromoacetamide to cause the production of 17a,21-dihydroxy-l6 a methylpr'egnane-ll1,2Q-trione-2l-acetate. Other known equivalent oxidizing agents such as those described hereinabove for preparing the 3,-keto derivative of the 16,6-

methyl compounds can, of course, also be employed. The

A -double bonds are then introduced into the A-ring by dihalogenation, e.g. dibromination of this 3-keto derivative by the rapid introduction of bromine in a suitable nonreactive organic solvent such as dioxane followed by dehydrohalogenation in a conventional manner. suchlas by refluxing the brominated steroid with dimethylformamide in the presence of calcium carbonate and lithium chloride, or with collidine or the like, or by use of other standard procedures as described hereinabove with relation to the preparation of the l6/3-methyldienes. The resultant product is lot-methylprednisone 2l-acetate.

Alternatively; 17a,21-dihydroXy lfiarmethylpregnane- 3,11,20-trione ZI-acetate can be modified by introduction solely of the A -double bond to form IGa-mBthylcOrtisone acetate. This is normally accomplished by halogenation of the former compound (preferably'wit'n bromine in a nisone Zl-acetate can then be saponified by reaction with conventional hydrolytic, reagents such as sodium or potassium bicarbonate as described hereinabove in connection with the corresponding 16,8-methyl steroids to remove the acyl-radical at C-21 and form the alcohol, 16a-methylcortisone, or 16a-methylprednisone. These compounds can be re esterified ifdesired to further enhance the duration of activity thereof as also described hereinabove with relation to the corresponding l6fl-methyl corticoids. The 16a-lower alkylsubstituted cortisone acylates such as the l6ot-methylcortisone acetate can also be converted to 16stmethylhydrocortisone by reaction thereof initially with semicarbazide, for example, to produce the 16,;x-methylcortisone acetate, 3,20-bis-semicarbazone, which is reacted with a metal borohydride reducing agent such as sodium or potassium borohydride to reduce the C-11 keto group to the ll-fl-hydroxy group While simultaneously saponifying the C-21 position to form 16u-methylhydrocortisone- 3,20-bis-semicarbazone. The latter product is then hydrolyzed by conventional procedures, e.g. dilute aqueous hydrochloric acid, to form 16oc-methylhydrocortisone.

This latter intermediate can then be transformed micrystallized out. In one preferred method, as described in Sequence E.

above, the'9ot halogen atom, and 'preferablythe 9a-tiuorine atom, is introduced into the 16a-rnethyl and l6fl-1nethyl 'methyl-M-pregnene-l7ot,2l-diol Zl-acetate. compounds are then reacted with hydrogen fluoride hydrocortisone acetate.

crobiologically according to procedures analogous to those described .in Belgian Patent No. 540,478 referred cto previously, hereinabove, employing a dehydrogenating microorganism, e.g. -Cryne5acterium simplex A .T;C.C. 6946), Bacillus sphaericus (A.T.C.C.' 7055), to form the'valuable anti-inflammatory steroid,

16o -methylprednisolone. The compound can then in 8 As noted earlier, the compounds of'our invention can be prepared by anumber of; alternative procedures. One

turn be re-esterified, as, for example, to form'an acylate with acetic anhydride in a suitable non-reactive organic solvent such as pyridine or lutidine and subsequently monoenes and dienes-21 esters as represented, for example, by lGa-methyl and 16,8-methyl prednisolone acetate or sulfonyl chloride, e.g. methane sulfonyl chloride, in alkaline organic media, for example, pyridine, 'to elfect the such as l6ot-methylprednisolone 2l-acetate. by reaction 1 the corresponding monoenes, loot-methyl or l6fl-rnethyl g hydrocor'tiso'ne, by dehydrating 1,60t-I'I16ii1yl or Me -methyl" prednisolone.acetate,',for example, with an alkyl or'aryl production of the corresponding A -derivative,'e.g.

Mot-methyl or 16t3-methyl-A -pregnatriene170L31- diol,-3,20-dione 21-acetate and 16a-methyl or lGB-methyl- A -pregnadiene 17 0:,21 diol 3,20-dione ZI-acetate. For conveniencein further describing this series of 'reactions, the o'tor ,8- isomerism of the16-rnethyl substituent will not normally be explicitly'referred to as in Sequence fEabove since this characteristic does not atiect the step- Wise reactions nor do these steps affect the specific 1600 or l6fi-isomeris'm of the reactant alkylated steroid. Placement .of a halogen, and preferably bromine at this coinplishedby conventionalfmeans, such as, for example, by reacting the A -steroid with hypobromous acid whichcan be and indeed is preferably prepared in situ from the reaction of, for example,N-bromoacetamide and perchloric acid yielding directly the corresponding .904-

- bromo-l6-rnethylpredniso1one 2l-acetate, and 9ct-bromo- '1 6-methylhydrocortisone ZI-acetate. types are useful anti-inflammatory compounds and are at Esters of j these the same time readily converted by oxidation to the corresponding ll-ketones by known techniques such as,,for example, with chromium trioxide in pyridine and readily hydrolyzed at the C-21 position by such standard hydrolytic' agents as aqueous methanolic potassium bicarbonate, sodium carbonate,. concentrated hydrochloric acid" in methanol-chloroform or like substances 'as disclosed hereinabove, to prepare the corresponding 21-alcohols.

Again alternatively, the.9ci-bromohydrins can be re fluxed with mild alkali, such as illustratively, sodium acetate in methanol, to form the corresponding 9,8,11 fi-oxido' "derivatives, 96,1113 oxido l6 methyl A -pregnadiene- 17ot-,21-diol-3,20-dione 21-,acetate, and 9,8,l1B-oxido-16- These latter in chloroform with or without ethanol and/or tetrahydrofuran to cause the: formation of 9ot-flu0ro-16-' methylprednisolone acetate and 9'ot-fluoro-l6-methyl- Similarly the substitution of point in the procedure at'the 9ot-carbon position is ac- I of these, by Way of illustration, results in theproduction of both the 16a-methyl and led-methyl steroids described herein and in which the 9a-halogen can be present in the starting steroid or added subsequently, by the procedures described immediately above. lllustrativebf these starting materials are 4,16-pregnadiene-21-ol-3,11,20-trione,

4,167-prgnadiene-1l;3,2l-diol-3,2t)-dione, and the 9a-halohydroxyl. The resultantproduct is reacted with pyruvic acid and p-toluenesulfonic acid to result-in the preparation of the corresponding l6-dehydro 20-keto-21-hydroxy-9sfluoro steroids, such as for example, the 9a-fiu0rO-4,16- pregnadiene-2l-ol-3,l 1,20-trione or 9a-fiuoro-4, leipregnadiene llfl,2l-diol-3,20-dione, referred to above. The reaction sequence for the preparationof the 16otmethyl and 16fl-methylsteroids of applicants herein from .9et-halogen and non-halogenated 20-keto-l6-pregnenes such as 4,16-

dienes will proceed in like manner by reaction thereof initially with nitroalkanes (e.g. nitromethane) in an organic base such as piperidineto form the corresponding 160t-I1l'fl0l'li6thYl derivative which can be reduced, e.g. with tin and hydrochloric acid to the l6cz-3IIli1lOII16ihYl steroid,

which, in turn, is quaternized with methyl iodide and subsequently pyrolyzed to form the corresponding l6-methylene derivative, which, in turn, converts to the 16-methyl- 20-keto-l6(l7 )-dehydro compound. Peroxidation of this latter product with hydrogen peroxide will cause the formation of the 16a,17a-oxido-l6fi-methyl-20-keto steroid which when reacted with hydrobromic acid, for example, will formthe corresponding bromohydrin which is debrominated to yield a mixture (separable by chromatography) of Ida-methyl and l6t3-methyl derivatives of, for

example, 4-pregnene-l7ot,2l-diol-3,ll,2 '-trione, 4-pregnone-11fi,17u,21-triol-3,20-dione and their 9oc-hal0 (e.g.

fiuoro) derivatives. These monoenes can be converted to the corresponding 1,4-dienes microbiologically employing, for example, the microorganism Bacillus sphaericus (A.'T.C.C. '7fi55)-of Coryneba c terium simplex (A.T.C.C.

'6946). ,These reaction steps are carried out by procedures well known to steroid chemists. It will, for example, be evident that, should'higher homologues of nitromethanes be employed (e.g. nitroethane, Z-nitropropane, nitrobutane and the like) on a 16-dehydrosteroid such as 3a-acetoxy-l6-pregnene-l1,20 dione, the corresponding alkyl homologues will result in the (3-16 position (eg.

16-ethyl- 1 6-dehydro 20-ketosteroids, lfi-isopropyll 6-de- 'hydro-Zflketosteroids, etc). The l6-alkyl-16-dehydro-20 Yketo steroids resulting from the pyrolysis of the quateranhydrous hydrogen chloride for hydrogen fiuoride'in 'this reaction results in the production ofthe corresponding 9 ct-chloro derivatives. ,Againthese compounds can beoxidized to the corresponding ll-lcetones by standard agents, e.g. chromium trioxide or N-halosuccinimides, or hydrolyzed to the corresponding 21-alcohols by standard saponification'procedures (e.g. aqueous methanolic potassium bicarbonate or concentratedhydrochloric acid in methanol). Thus the preferred halogenated compounds of our invention, 9ot-fluoro-l6-lower alky'l prednisolones and 9a-fluoro l6-lower alkyl prednisones and particularly .9a-fluoro-l 6a-methylprednisolone, v prednisone', .9a-1iuoro-16 3-methylprednisolone and 9a-tluoro-'16p3-methylprednisone are prepared by these procedures.

nized steroids can be converted to corticoidsiin the manner already described, e.g;, hydrogenation of the double 1 bond with palladium on carbon orRaney nickel,- introduction of the 17-hydroxy group by meansof enol acetylation and peroxidation (e.g., acetic anhydride andp-toluenesulfonic acid followed by treatment with perace'tic acid), introduction of the 21-acetate by meansof bromination and acetoxyl ation (sodium acetate in dimethylformamide) and elaboration of the A ring in the conventional manner to introduce the A or o -double bond(s) and 3-lreto group.

Alternatively, they may be epoxidized with alkaline hydrogen peroxide to yield 16a,l7a-epoxy 1fi-alkyl-ZG-lceto 7 These are then treatedwith a hydrohalic acid suchas hydrogen bromide and debrominated, e.g., with Zinc dust or Raney nickel or palladium and hydrogen. This results in the formation of both l6a-alkyl-17w-hy i droxy-ZG-keto steroids, and lGfi-alkyi-lM-hy ,oxyQO- V steroids.

' l6a-methylprednisone 2l-acetate.

l'reto steroids. After separation, e.g., by chromatography, these are then handled in the usual fashion, e.g.,the 21- acetate is introduced via bromination and acetoxylation, and the A ring elaborated via oxidation of the 3-hydroxyl (if present), followed by bromination-debromination.

Since the higher diazoalkanes are rather difficult to prepare, it is preferable to convert 160t-filkYl-20-k6t0 steroids to 16B-alkyl-20-keto steroids by a relatively simple procedure. The 16a-alkyl steroids are brominated at position 17, e.g., with bromine in an inert solvent (e.g. pyridine), or with N-bromosuccinimide, and the resulting compound dehydrobrominated, e.g., with collidine or with dimethylformamide to give 16-alkyl-l6-dehydro-20-keto steroids. Hydrogenation of these gives 16B-alkyl-20-keto steroids.

Thus far, we have described as starting materials only compounds which are available from bile acids; There are, however, several other classes of starting material which are extremely useful, e.g. diosgenin and smilagenin. For example, diosgenin is readily converted to l6-dehydropregnenolone which can be converted readily to l6et-methylpregnenolone by means of the Grignard reaction and to l6-methyl-16-dehydropregnenolone by means of the di azomethane procedure. The former compound is hydrogenated, e.g. by means of palladium catalyst, to 16amethylallopregnane-3p-ol-20-one and the cortical side chain elaborated in the usual way (enol acetylation at C17 and epoxidation to give the 17a-hydroxy-20-ketone followed by bromination and acetoxylation at (3-21). The C-3 hydroxyl is oxidized to the ketone, a 2,4-dibromide is prepared using bromine and dioxane or other inert solvent and dehydrobromination is effected by means of collidine or dimethylformamide to yield 16a-methyl- A -pregnadiene-l7a,2l-diol-3,20-dione 21-acetate. This can be hydroxylated at position C-ll to yield either 16amethylprednisolone or l6a-methyl-ll-epi-prednisolone. This last compound may be acetylated selectively at C-21 to yield a mono-acetate and then oxidized at C-ll to give Also, l6a-methyl-l.1- epiprednisolone 21acetate may be dehydrated at (3-11 by tosylation of the c-llot-hydroxyl followed by treatment with sodium acetate to yield l6a-methyl-A -pregnatriene-17a,21-diol-3,20-dione 2l-acetate, a compound previously described as an intermediate for the preparation of 9e-halo-l6a-metl1ylcorticoids.

Alternatively, 160% or 16B-alkyl-allopregnane-l7a,2l-

diol-3,20-dione or its 2l-acetate can be hydroxylated in the ll-position, e.g. with Rhizopus nigricans (A.T.C.C.

6227) or Curvularia lunata (N.R.R.L 2380), the C-21 hydroxyl is acetylated and the 3-keto-A or 3-keto-A system introduced in the usual manner.

The 16 methyl 16 dehydropregnenolone described above is hydrogenated to l6 8-methylallopregnane-36-01- 20-one. This is treated in an exactly analogous fashion to yield 16fl-methyl-A -pregnadiene 170:,21 did-3,20-

dione 2l-acetate, 16/3-methylprednisolone 2l-acetate, 16B- methyl-ll-epi-prednisolone 2l-acetate, and -16,B-methyl- A -pregnatriene-l7oc,21-diol-3,20-dione Zl-acetate (a compound previously described as an intermediate for the preparation of 9ahalo-16,6-methyl corticoids).

Other important starting materials are the 12-oxygenated sapogenins, such as hecogenin, roclrogenin and gentrogenin. For example, hecogenin has been converted to ll-keto-16-dehydroallopregnane-3,8-ol-20-one. A 16stmethyl substituent can be introduced by means of the Grignard reaction and the side chain and A ring elaborated in the usual manner to contain the A or A -double bond(s) and 3-keto and 21-acyloxy or hydroxyl groups, or the l6/3-methyl substituent can be introduced via the diazomethane reaction and the side chain and A ring elaborated again by these same procedures.

Certain of the novel compounds of our invention, that is, the l6ccand 16fl-alkyl-substituted derivatives of cortisone, hydrocortisone, prednisone and prednisolone, nonhalogenated or halogenated in the 9a-position, as well as their C-21 esters possess, as has been noted above, valuable anti-inflammatory and diuretic (i.e., sodium and water excretory) propenties (thus explicitly excepting applicants novel 11oc-hydroxy-l6-alkylated steroids, which are, however, valuable as intermediates in the formation of the corresponding ll-keto steroids, e.g., l6-alkyl cortisone and l6-alkyl prednisone). The alkylated (notably the 16et-methyl and 16B-methyl-substituted) dienes with or without the 9a-halogen substituted are particularly valuable therapeutic agents possessing similar therapeutic effects in the treatment of inflammatory diseases, such as for example, arthritis, dermatitis, asthma, and the like, and are normally administered in a daily maintenance dosage range of 0.50 to 10 mg. the preferred daily dosage of the non-halogenated dienes being from about'2 mg. to about 10 mg. and that of the halogenated, and preferably the'9a-fiuoro-alkylated dienes being in the range of 0.5 mg. to 5.0 mg.

.The non-halogenated l6a-alkyl and l6 8-alkyl monoenes of our invention, are, in addition to being valuable intermediates in the formation of the corresponding dienes, also active as anti-inflammatory agents and in inhibiting renal tubular reabsorption of sodium. A daily dosage of 25 mg. to mg. is recommended for oral administration of these monoenes to induce sodium and water. diuresis, e.g. in the treatment of ascites, congestive heart failure and cyclical edema.

Similarly, the 9-hal0-l6ozand 9-halo-l6/3-alkylated monoenes of' our invention are also useful as antiinfiammatory agents, since they manifest an enhanced activity in the treatment of such inflammatory diseases as arthritis, dermatitis, asthma, and the like. When ad ministered orally, the recommended daily dosage is in the range of 4 mg. to 15 mg. When treatment so indicates, these or 16B-alkylated dienes and monoenes can be administered parenterally in the form of therapeutically acceptable solutions and suspensions, e.g., in aqueous media, or where oral administration is indicated, can be incorporated into tablets (normally from about 0.50 mg. to 5 mg. per tablet), elixirs and other known pharmaceutical dosage forms by standard procedures. They can also be administered topically in the form of ointments or creams, or as solutions, e.g. in dimethyl acetamide or diethyl acetamide, or in the form of suppositories dissolved or suspended in a fatty or waxy vehicle which melts at approximately body temperature,'or as aerosols when mixed with suitable materials, such as isopropylmyristate and dichloro difiuoromethane (Freon).

The following examples are further illustrative of the invention Example 1 1619-METHYLCORTISONE 21-ACETATE N, 6.76. Found: C, 69.51; H, 7'.98; N, 6.69.

(B) l6-methyl 16 pregnene-3u-0l-1I,ZO-dione 3-acetate.The pyrazoline of Example 1A (3.00 g.) is heated under reduced pressure to about 210 C. until the evolution of nitrogen ceases. The resulting oil is cooled to room temperature, then crystallized by the addition of ether. The precipitate is filtered and dried, yielding 2.12 g. of 16-methyl-16-pregnene-3ot-ol-l1,20-dione 3-acetate, lvLP. 163-166 C., [eth l-69.9 (dioxane) Amax. 248 ma (6 10,800).

' ll AnalysisrCalcd. for C H O C, 74.57; H, 8.87. Found: C, 74.85; H, 8.55.

A solution of 12.0 gi of the 16-pregnene compound of Example 1B in 250 ml. of glacial acetic acid is hydrogenated at room temperature and atmospheric pressure in the presence of 3.0 g. of palladium on charcoal catalyst. After the reduction is completed (about 1.5 hours), the catalyst is removed by filtration, and the filtrate concentratedunder reduced pressure to about 100 ml. The residue is poured intowater, and the precipitated solid filtered, washed and dried, yielding 11.4 g. of impure lee-methylpregnane-Sa-ol-l1,20=dione 3-acetate, Ml. 148-155 C. After recrystallization from acetone-hexane there is obtained8.40 g. of the product of this Example 1C, MP.

1*60-163 C., no U.V. absorption at 220-300 mu,

Analysis..-Calcd. for C l-1 0 C, 74.19; H, 9.34. Found: C, 74.37; H, 9.06.

(D) 16B methylpregnane 3a,]7or-di0l-11,20-cliOn.- A solution of 6.77 g. of 16/3-I1'l6thYlPl6gl'l3Il6-30t-Ol-11,20- dione 3-acetate, in 156 ml. of acetic anhydride containing 3.89 g. of p-toluene-sulfonic acid is kept at 100 C. for six hours; during which time about 16 ml. of distillate is removed every half hour by the application of vacuum. The resulting oily residue is dissolved in'80 ml. of benzone and washedthree times with water, then with a solution of 1.55 g. of sodium acetate in 20 mlfof water, The benzene layer is dried over magnesium sulfate, and then stirred for 18 hours at C. with a mixture of 0.52 g. of sodium acetate in 12 ml. of commercial peracetic acid. Excess peracetic acid is then destroyed by the drop- Wise addition of a solution of 15.5 g. of sodium sulfite in 52 ml. of water, while maintaining the temperature between 10? C. and 20 C. An additional 1.57 g. of sodium sulfite is then added, and the mixture stirred overnight until a starch-iodide test is negative. The benzene layer is separated, washed three times with water, and evaporated. To the resulting residue which is dissolved in 345 ml. of methanol, there is added a solution of 3.62 g. of

sodium hydroxide in 39.5 ml. of water, and the mixture refluxed for 15 minutes, After neutralization with 4' ml. of glacial acetic acid, the solution is concentratedunder reduced pressure to a volume of about ml. This concentrate is poured into. a mixture of ice and water and the solid which precipitates is filtered and dried,

yielding 6.69 g. of impure 16,Q-rnethylpregnane-3a,17a-

diol-11,20-dione, M.P. 105140 C. This is chromatographed on Florisil, and the material eluted with 3350% ether hexane, is combined and crystallized from acetonehexane, yielding 2.09 g. of 16B-methylpregnane-3oc,170tane).

Analysis.-Calcd. for C H O C, 72.89; H, 9.45. FOlllldZzC, 72.82; H. 8,25.

165 methylpregnane-3ot,17zx-diol-11,20-dione 3-acetate ,isprepared by adding to 1 gram of the corresponding 16,8-

methylpregnane-diol a solution of 0.5 ml. of acetic anhydride in 2.3 ml. of pyridine. After standing one hour 21-acerate.--A solution of 362 mg. of 16/3-methylpregnane-3ct,17u-diol-11,20-dione in 18 m1. of chemically pure (C.P.) chloroform (containing afew drops of chloroform previously saturated with hydrogen bromide) is brominated at 25 C. by the addition (over a three-hour period) of 165 mg. of bromine in 10 ml. of chloroform.

After removal of the solvent under reduced pressure,

10ml. of dimethylformamide and one gram of sodium acetate are added. The mixture is stirred at 60 C. for two hours, allowed to stand overnight at room temperature, then heated another two hours at 60 C. The mixture is then poured into Water, and the precipitated solid filtered and dried, yielding 370mg. of impure 1 613-methylpreg nane-3a,17a,21-triol-11,20-dione 21-acetate, 'M.P. 192- 203 C. Recrystallization from acetone hexane gives 280 mg. of the product of this example, M.P. 99205 C. The analytical sample, after another recrystallization, melts at 200-2055" C.

Analysis.-Calcd.' for C H O C, 68.54; H, 8.63;

Found: C, 68.79; H.839.

(F) 16B-mezhylpregnane-170,2l-di0la3,11,20-tri0ne 21- acetatd-A solution of 180 mg. of 16fi-methylpregnane- 3a,l7cc,2l-tIl0l-11,20-dl0n6 21-acetate, in 30 ml. of acetone-Water is cooled to 10 C. One drop of concentrated hydrochloric acid is added, along with 150 mg. ofN-brornosuccinimide, and the mixture allowed to react 22 hours at 10 C. in the dark. Excess sodium sulfite solution is then added, and the mixture concentrated under reduced pressure to a small volume from which precipitates 120 mg. of impure 16,8-methylpregnane-17a,21- diol-3,11,20-trione 21-acetate, M.P. 19 0-195 C. dec. Purification is effected by means of zinc dust in aqueous acetone containing 2 drops of acetic acid. The zinc is removed by filtration, the filtrate concentrated, and hot solution is evaporated to dryness under reduced pressure.

The residue is slurrie'd with water, filtered and dried to yield mg. of 4o-bromo-16,13-methylpregnane-17a,21- diol-3,l1,20-trione 2l-acetate Ml. 115-130 C. dec., which is of sufficient purity for the conversion described in the following procedure.

(H) 16,8-methylcortisone 21-acetate.--The 4fi-bromolefl-methylpregnane-17a,2'1-diol-3,11,20-trione 21-acetate of above Example 16 is dissolved in a mixture of 30 mg. of semicarbazide, 4 ml. of tert.-butyl alcohol and 2 ml. of methylene chloride, and stirred in a nitrogen atmosphere for two hours at 25-30 C. After removing the solvent under reduced pressure, the residue is dis solved in 5 ml. of 80% acetic acid-water along with excess 70% pyruvic acid, and the reaction mixture allowed to stand at room temperature for 20 hours. The solution is brought to about pH 7 with dilute (5%) sodium hydroxide, and then is extracted with methylene chloride.

The organic extracts are evaporated to a residue, and the residue chromatographed on Florisil. Chrystalline material (25 mg.) obtained from the 50% ether-hexane and ether eluates, on crystallization from acetonehexane yields 20 mg. of 16B-methylcortisone 21-acetate,

M.P. 198208 G. Mfg? 238 m (e 12,800)

' Example 2 16,6-METHYLCORTISONE One gram of lo e-methylcortisone ZI-acetate (prepared as in Example 1) is dissolved in 25 ml. of methanol and 5 ml. of water containing 0.2 g. of posassium bicarbonate. This solution is refluxed for 6. hour, then concentrated under reduced pressure. Water is added to the residue, and the resulting precipitate is filtered and dried. Crystallization from acetone-hexane gives 16,8-methylcortisone.

example 3 lGB-L'IETHYLHYDROCORTISONE (A) 16B-methylc0rtisoi1e3,20 bis semicarbazonar- A mixture of 5.0 g. of 16fi-methylcortisone,8.3 g; of semi-' 13 carbazide hydrochloride, 6 g. of pyridine, 50 ml. of water and 200 ml. of methanol is refluxed for 16 hours. The solution is concentrated to about 60 ml., then poured into water to precipitate l6,6-methylcortisone-3,20-bis-semicarbazone.

(B) 165 methylhydrocortisone 3,20 bis semicarbazone-A solution of 6.0 g. of the bis-semicarbazone of above Example 3A and 4 g. of potassium borohydride in 200 ml. of tetrahydrofuran and 100 ml. of water is refluxed for 6 hours. The solution is cooled, and acetic acid added to pH 5.5. The organic solvent is distilled and the solids in the residue are filtered to give 16B-methylhydrocortisone 3,20-bis-semicarbazone.

(C) 16fl-methylhydrocortisone.Under a nitrogen atmosphere, 5.0 g. of the bis-semicarbazone of above Example 3B are dissolved in 250 ml. of 2.4 N hydrochloric acid. The solution is cooled to C., then 2.5 g. of sodium nitrite in 25 ml. of water is added over a minute period at 5 C. The reaction mixture is stirred an additional 30 minutes, then cooled to below 15 C., neutralized with sodium hydroxide, and extracted several times with chloroform. The solvent is evaporated under reduced pressure to give a solid residue which, after crystallization from acetone-hexane, yields 16 3-methylhydrocortisone.

Example 4 lfifl-DIETHYLHYDROCORTISONE ZI-ACETATE To one gram of 16B-methylhydrocortisone, prepared as in Example 3, is added 0.5 ml. of acetic anhydride in 2.3 m1. of pyridine. After standing for one hour at room temperature, the mixture is poured into ice and hydrochloric acid. The resulting precipitate is filtered and recrystallized from aqueous methanol to yield 16/3-methylhydrocortisone 21-acetate.

Example 5 IBa-METHYLCORTISONE 21-ACETATE (A) 16a-methylpregnane-3a-0l-11,20-di0ne.A mixture of 3.72 g. of 16-pregnene-3a-ol-11, 20-dione in 20 ml. of dry toluene is added to a Grignard reagent prepared from 7 g. of methyl iodide and 1.2 g. of magnesium in 40 ml. of ether, and containing 200 mg. of cupric chloride. The reaction mixture is distilled until a vapor temperature of 100 C. is reached. The distillation is then stopped, and the reaction temperature maintained at 100 C. for five hours. The mixture is then cooled, poured onto ice and an aqueous solution of ammonium chloride, and the solvent layers separated. The organic layer is distilled in vacuo to an oily residue (398 g.) which is chromatographed on Florisil. The 20% ether-hexane eluates are combined and evaporated to give 2.08 g. of a residue which is crystallized from acetone-hexane to yield 16a-methylpregnane 3a ol 11,20 dione, M.P. 149-154 C., [u] +100.5 (dioxane).

(B) 160: methylpregnane 3a,]7u-di0Z-11,20dione. 16ct-methylpregnane-3a-ol-11,20-dione (7.44 g.), prepared as in above Example 5A is dissolved in 156 ml. of acetic anhydride containing 3.89 g. or p-toluenesulfonic acid. The reaction mixture is heated to 100 C., and about 10 ml. of distillate removed in vacuo every minutes. After 6 hours, the remaining oily residue is dissolved in 80 ml. of benzene, and the solution washed three times with water, and once with a solution of 1.55 g. of sodium acetate in 20 ml. of water. A mixture of 12 ml. of 40% peracetic acid containing 0.52 g. of sodium acetate is added to the benzene solution and the mixture stirred at room temperature for 19 hours. A solution of 15.5 g. of sodium sulfite in 80 ml. of water is added at a temperature below 25 C., then an additional 1.6 g. of sodium sulfite is added and the mixture stirred overnight. The solvent layers are separated. The organic layer is washed three times with water, dried over calcium chloride, filtered, and evaporated to an oily residue under reduced pressure. The residual oil is dissolved in 345 ml. of methanol and 39.5 ml. of

water containing 3.62 g. of sodium hydroxide, and the which is chromatograph'ed on Florisil.

mixture refluxed 15 minutes. After neutralizing the excess alkali with 5 ml. of acetic acid, the methanol is distilled in vacuo. Water is added to the resultant residue, and a solid (5.25 g., M.P. 173179 C.) separates which, when filtered and crystallized from acetone, yields 2.36 g. of 160: methyl-pregnane-30:,17ot-di0l-11,20-dione, M.P. 18l.5184 C., [u] +74.4 (dioxane).

t (C) 1604 methylpregnane-3a,17a,21-tri0l-11,20-di011e 21-acetate.A solution of 362 mgm. of the l6a-methylpregnane-diol of Example 53 in 18 ml. of chloroform is brominated at 20 to 30 C. by the dropwise addition of 160 mg. of bromine in 10 ml. of chloroform over a 45 minute period. Stirring is continued an additional 20 minutes, then the solvent removed under reduced pressure at 10 C. The residue is dissolved in 10 ml. of dimethylformamide, one gram of sodium acetate added, and the reaction mixture stirred at 60 C. for 12 hours. The reaction mixture is then poured into 10 ml. of concentrated hydrochloric acid and ice, and the precipitated solid filtered, washed with water and dried: 250 mg, M.P. 150- 158. The filtrate is extracted with methylene chloride and the extracts evaporated to give an oily residue of 180 mg. The solid (250 mg.) and the oil (180 mg.) are com bined and chromatograped on Florisil. Eluates of 40% ether-hexane are combined and distilled to a residue mg, M.P. -155 C.) substantially of 16a-methylpregname-3a,17a,2l-triol-11,20-dione 21-acetate. This product is used without further purification as the starting compound in the procedure immediately following.

(D) 160: methylpregnane 1701,21-di0l-3,11,20-trione 21-acetate.-To a solution of 110 mg. of the product of above Example SC in 2.5 ml. of 80% tert.-butyl alcohol, 1.0 ml. of glacial acetic acid and one drop of concentrated hydrochloric acid is added 100 mg. of N-bromoacetamide at 5 C., and the reaction mixture kept at 5 C. for 20 hours. Aqueous sodium bisulfite solution is added, and the mixture extracted with methylene chloride. The 0- ganic solvent extracts are evaporated to give a residue which is debrominated by treatment with zinc dust in acetone-acetic acid. After filtering the zinc from the reaction mixture, water is added to the filtrate. The resultant precipitate is filtered and dried to yield 40 mg. of 16a-methylpregnane-17a,21-diol-3,11,20-trione 21-acetate, M.P. 205211 C.

(E) 4 bromo 16a methylpregnane 1704,21 diol- 3,1],20 trione 21-acetate.-16a-methylpregnane-17a,21-

diol-3,11, 20-trione (80 mg), prepared as described in' Example 5D is dissolved in 3 ml. of tert.-butyl alcohol and 3 ml. of methylene chloride and is brominated at 3035 C. by the rapid addition of a solution of 32 mg. of bromine in 4 ml. of tert.-b utyl alcohol. Bromination is complete in 2 hours, and the solution evaporated to dryness under reduced pressure. The resultant residue is slurried with water, filtered and dried to give 92 mg. of a solid of substantially 4-bromo-16a-methylpregnane-17a,21-diol, 3,11, 20-trione 21-acetate, which is used without further purification in the procedure immediately following.

(F) 160; methylcortisone 21 acetate.4-bromo-16amethylpregnane-17a,21-diol-3,11,20-trione 21-acetate (92 mg.) of above Example SE is dissolved in a mixture of 30 mg. of semicarbazide, 4 ml. of tert.-butyl alcohol and 2 ml. of methylene chloride, and stirred in a nitrogen atmosphere for two hours at 2530 C. The solvent is removed in vacuo and the resultant residue dissolved in 5 ml. of 80% acetic acid-water, along with an 8 molar excess of 70% pyruvic acid with respect to the molar quantity of the 4-bromo-16a-methylpregnane starting compound. The reaction mixture is allowed to stand at room temperature for 20 hours. Dilute (5%) aqueous sodium hydroxide is added until the. reaction mixture is at about pH 7, then the solution is extracted with methylene chloride. The organic extracts are evaporated to a residue The 50% etherhexane eluates and the 100% ether eluates are combined and evaporated to a crystalline residue (25 mg.) which is 15 crystallized from acetone-hexane to give-20 mg. of 160'.- methylcortisone 21-acetate,

XQESQ 238, III/.5 (6 14,800)

. Example 6 mwMETHYLCORTISONE 21-ACETATE 16a-methylcortisone 21-acetate (0.5 g.), prepared as in Example 5, is dissolved in 20 m1. of methanol and 2 ml. of water containing 0.1 g. of potassium bicarbonate. The solution is refluxed for A2 hour, then concentrated to /2 the volume in vacuo, and water added. The precipitated solid is filtered, dried, and crystallized from methanol to yield 16a-methylcortisone,

A235,? 238 III .5 (6 15,200) Example 7 1fia-METHYLHYDROCORTISONE- (A) 16a-methylc0rtis0ne ZJ-acetate 3,20-bis-semicarbazne.A mixture of l6a-methylcortisone 21aacetate (2.5 g.), prepared as in Example 4.2 g. of semicarbazide hydrochloride, 3 g. of pyridine, 25 ml. of water and 100 ml. of methanol is refluxed for 16 hours. The solution is concentrated to about 30 ml., then poured into water to precipitate an almost quantitative yield of 16a-methylcortisone 21-acetate 3,20-bis-semicarbazone which is filtered and dried, and used without further purification in the procedure immediately following. i

(B) 160: methylhydrocortisone 3,20-bis-semicarbazone.A solution of 3.0 g. of the bi-s-semicarbazone of Example 7A and 2 g. of potassium borohydride in 100 ml. of tetrahydrofuran and 50 ml. of water is refluxed for 3 hours. The solution is cooled, and acetic acid added to pH 5.5. The organic solvent is removed by distillation, and the solids collected by filtration to give 16a-methylhydrocortisone 3,20-bis-semicarbazone which isused with- V out further purification in the procedure immediately following.

(C) 1(Soc-methylhydrocortisone.--Under a nitrogen atmosphere, 5.0 g. of the bis semicarbazone of Example 7B is dissolved in 250 ml. of 2.4 N hydrochloric acid. The solution is cooled to 5 C., and a solution of 2.5 g. of sodium nitrite in 25 m1. of Water is added at 5 C. over a minute period. The reaction mixture is stirred an additional 30 minutes, and then is brought to neutrality with sodium hydroxide, at a temperature below 15 C. The reaction mixture is extracted several times with chloroform. The chloroform extracts are combined and distilled in vacuo to a solid residue which, when crystallized from acetone yields 16a-methylhydrocortisone,

x3 52 240 mu (6 14,700) Example 8 1Ga-METHYLHYDROCORTISONE 21-ACETATE 16a-methylhydrocortisone (100 mg.), prepared as in Example 7, is dissolved in 2 ml. of pyridine containing 100 mg. of acetic anhydride and is allowed to stand at room temperature for one hour. The reaction mixture is poured into ice and hydrochloric acid, and a solid precipitates which is filtered and crystallized from aqueous methanol to yield 16a-methylhydrocortisone 21-acetate, AXffi 240 mu (615,000) Example 9 lfia-n-BU'IYLCORTISONE ZI-ACETATE (A) 16a n-butylpregnane 3a-0l-11,20-di0ne.-16apregnene-3a-ol-l1,20-dione is reacted with n-butyl-iodide and magnesium in the manner of Example 5A to yield 16a-n-butylpregnane-3a-ol-l1,20-dione, which is crystallized from acetone-hexane.

(B) 16a n-butylpregnane-3a,17a-di0l-11,20-di0ne.- 16a-n-butylpregnane-3a-ol-l1,20-dione, prepared as in Example 9A, is first reacted with p-toluenesulfonic acid in acetic anhydride, then with 40% peracetic acid and sodium acetate, and finally with aqueous sodium hy- 1 droxide inthe manner described in Example 53. A solid product is obtained which, when crystallized from acetone yields 16a-n-butyl-pregnane-3a,17a-diol-11,20-dione.

(C) 16a n-butylpregnane3a,17a,21-tri0l-11,20-di0ae 21 acetate.'16a-n-butylpregnane-3a,l7a-di0l-11,20-dione, prepared as in Example 9B, is brominated and then treated with sodium acetate in dimethylformamide in the manner of Example 50. The reaction product is isolated and purified in the manner described to give 16a-n-butylpregnane-3a,17a,21-.triol-11,20-dione 21+acetate.

(D) 160: n batylpregnane-I7a,2]-di0l-3,11,ZO-trione 21 acetate.16a-n-butylpregnane-3 ,17a,21-trio1-11,20- dione 21-acetate, prepared as in Example 9C, is reacted with N-bromoacetamide, and the resultant product isolated and purified in the manner of Example 5D to yield 16a n-butylpregnane-l7a,21-diol-3,11,20-trione 21-acetate.

(E) 4-br0m0-16a-nbutylpregnane-] 7a,21-diOl-3,11,Z0 trion-e 21 acetate.16a n-butylpregnane-17a,21-diol- 3,11,20-trione 21-acetate, prepared as in Example 9D, is brominated and the product isolated in the manner of Example SE, to give 4-bromo-16a-n-butylpregnane-17a,21- diol-3,11,20-trione 21-acetate which is used without purification in the following reaction.

(F) 16a-n-batylc0rtis0ne 21 -acetate.-The 4-bromopregnane, prepared in above Example 9E, is reacted with semicarbazide, then with acetic acid and pyruvic acid in the manner described in Example SF. The product is isolated and purified in the described manner to yield 16a-n-butylcortisone 21-acetate,

' kfiiff 238 m l (e14,800)

Example 10 IGa-n-BUTYLCORTISONE 16a-n-butylcortisone 21-acetate, prepared as in Example 9, is hydrolyzed with potassium bicarbonate in methanol and water in the manner of Example 6 to give l6a-n-butylcortisone,

, Example 11 10a-n-BUTYLCORTISONE 2l-n-BU'1YRATE 16a-n-butylcortisone mg.) prepared as in Example 10, is dissolved in 2 ml. of pyridine containing 100 mg. of butyric anhydride, and left at room temperature for one hour. The reaction mixture is then poured into ice-hydrochloric acid. A precipitate forms which is filtered and crystallized from aqueous methanol to give 16a-n-butylcontisone 2 l-n-butyrate;

Am? 238 m (614,500)

Example 12 (A) 16,8 methyl-4,9-pregnadiene-Z7u,21-di0l-3,20-dione 21 -acetate.To a solution of 16,8-methylhydrocortisone 21-acetate (0.3 g.), prepared as in Example 4, in 5 ml. of pyridine there is added 0.2 ml. of benzenesulfonyl chloride in 3 ml. of pyridine. The solution is allowed to stand for 4- hours, then is poured into ice-hydrochloric acid. A solid precipitate which is. filtered and crystallized from acetone-hexane to give 16,8-methyl-4,9- regnadicne- 17a,2l-diOl-3,20-di01'16 2l-acetate,

(B) 9a-br0m0-16,8-methylhydrocortisone 21-acetate.

-A suspension of 0.2 got the 16/3-methyl-4,9 pregnadi- A mixture of 0.5 g. of 9a-bromo-16fi-methylhydrocortisone ZI-acetate, prepared as in Example 12 in 100 m1. of methanol, 20 ml. of chloroform, H11. of water and 5 ml. of concentrated hydrochloric acid is allowed to stand 48 hours at room temperature. Water is then added, and the mixture extracted with methylene chloride. The organic extracts are washed With'water, dried and concentrated to a residue. Crystallization of this residue from acetone yields 9a-bromo-l6B-methylhydro cortisone. t

. Example 14 9a-FLUO RO-1Gfl-lVIETHYLHYDROCORTISONE 21-ACE'IATE (A) 95,113 oxido 16,8 methyl 4 pregnene 170:, 21 diol 3,20 dione 21 acetate.To 9 u-bromo-l6fimethylhyd-rocortisone' 21-acetate (0.3 g.), prepared as in Example 12, in 20 ml. of methanol thereis added 0.3 g. of potassium acetate. The mixture is refluxed for 2 hours, then concentrated in vacuo to a residue. Water is added to the residue, and a. solid separates which is filtered and crystallized from methanol-water to give 0.1 g. of 9,8,115- oxido 16B methyl 4 pregnene 17a,2l. diol 3,20- dione 21 acetate.

(B) 911 fluoro 16,8 methylhydrocortisane 21 acetate.-A solution 0.2 g. of the 9B,11/i-oxido-4-pregnene of above Example 14A in 10 ml. of alcohol-free chloroform is saturated with anhydrous hydrogen fluoride at 0 C. The mixture is allowed to stand 4 hours at 0 C., then concentrated to a residue in vacuo. Crystallization of the residue from acetone-hexane gives 0.1 g. 9u-fiuoro l6/i-methylhydrocortisone 21-aoetate.

Example Qa-CHLORO-lGB-METHYLHYDROCORTISONE 21-ACETATE A solution of 0.2 g. of the 9,8,11B-oxido-4-pregnene of Example 14A in 30 ml. of alcohol-free chloroform is saturated at 0 C. with anhydrous hydrogen chloride, and the mixture allowed to stand at 0 C. for six hours. The

solvent is distilled in vacuo from the reaction mixture leav- In the manner described in Example 13, 9oz-fll10fO-l613- rnethylhydrocortisone 2l-acetate, prepared as in Example 14, is converted to 9a-fiuoro-lfifl-methylhydrocortisone by means of hydrochloric acid in methanol-chloroform-water.

Example 17' A solution of 0.2 g. of sodium sulfite in 4 ml. of

water is added, then the solution is extracted with methylene chloride. The organic extracts are washed with water, dried, filtered and evaporated to a residue which is crystallized from methanol to yield 0.1 g. of 9a-bromo-16amethylhydrocortisone 21-acetate,

$3.? 242 ml. (e16,400)

Example 18 9a-FLUORO-lSa-METHYLHYDROCORTISONE (A) 95,11 3 oxido 16o; methyl 4 pregnene 17a, 21 diol 3,20 dz'one 21 acetate.A mixture of 0.3 .g. of 9a-bromo-l6a-methylhydrocortisone 2l-acetate, prepared as in Example 17, in 20 ml. of methanol and 0.3 g. of potassium acetate is refluxed for 2 hours, then concentrated to a residue. Water is added to the residue, and a solid separates which is filtered and crystallized from methanol to give 0.1 g. of 9 3,1lfi-oxido l6a-methyl-4- pregnene-l7a,2l-diol 3,20-dione ZI-acetate.

(B) 911 fluoro 16oz methylhydrocortisone 21 acelate.A solution of 0.2 g. of the 913,1 1fi-oxido-16a-methyl- I 4-pregnene of above Example 18A in 10 ml. of alcoholsa-nnorro41ea rrnrnrnrrynnoconrrsonn 'z'l-aontratrn (A) 16 methyl- 4.9- pregnadieae 1 70.,21 diol- 3, 20 dione 21 acetate.l6oc -''-methylhydrocortisone 2lacetate (0.2 g.), prepared as in Example 8, is dissolved 3 ml. of pyridine and 0.1 ml. of benzenesulfonyl chloride in 2 ml. of pyridine is added. The solution is allowed to stand for 4 hours, then is poured into ice-hydrochloric acid. 'A solid precipitates which is filteredand "crystallized from acetone to give l6 -methyl-4,9-pregnadienel7a,2l -diol-3,20-dione 2l-acetate, r

n 233 m teleeooi" max.

hours.

free chloroform is saturated with anhydrous hydrogen fluoride at 0 C. The mixture is allowed to stand 5 hours at 0 C., then concentrated to a residue under reduced pressure. Crystallization of the residue from acetonehexane yields 0.1 g. of 9a-fluoro-16a-methylhydrocortisone 2l-acetate,

mg? 238 m (616,200)

(C) 9a flaoro 16cc methylhydr0c0rtis0ne.1n the manner described in Example 13, the ZI-acetate ester prepared in Example 18B, is converted to the free alcohol, 9a-fiuoro-16a-methylhydrocortisone, by means of hydrochloric acid in methanol-chloroform-water.

, g Example 19 IGB-IkBUTYLCORTISONE ZI-ACETA'IE (A) 17 bromo 16oz n butylpregnarze 3a 0l 11, 20 dione.'One gram of 16a-n-butylpregnane-3a-ol-l1, 20-dione, prepared as in Example 9A, is dissolvedin 10 ml. of acetic acid and is then brominated with 1.1 equivalents of bromine in acetic acid. When the bromine color is discharged, the solution is poured into water and there is precipitated a solid of substantially 17-b1'0II10-l6a-I1- butylpregnane-3a-ol-l1,20-dione. This l7-bromo-prodnot is filtered and used in the next procedure without further purification. V

(B) 16 n butyl #16 pregnene 3a ol 11,20-di- 0ne.To the l7-bromo-l6a-n-butylpregnane prepared in above Example 19A there is added 10 ml. of dimethylformamide, and the reaction mixture is refluxed two hours.-

The solution is poured into dilute hydrochloric acid and fate, filtered, and evaporated in vacuo to aresidue of sub stantially l-n butyl-16rpregnene-3a-ol-ll,20 dione which is used without further purification in the reaction procedure immediately following. i i (C) n.- butylpregnane 3a ol 11,20 di0ne. The 16-n-butyl-l6-pregnene-3ran-01 11,20-dione, prepared in above Example 193, is dissolved in 10 ml. of acetic acid and hydrogenated using palladium on charcoal as catalyst. When one equivalentof hydrogen'is absorbed, the reaction i-sstopped, the catalyst removedby filtration, and the filtrate poured. into water. A solid precipitates which is filtered and crystallizedfrom acetone-hexane to give16fi-n-butylpregnane 3wolel1,20-dionei (D) 16 8 n butylpregaane L 3a,17a diol 11,20 dione .In the manner-described in, Example 1D, 16/8-nanhy'dride, then with sodium acetate 'in 40%"peracetic acid, and finally withaque'ous sodium hydroxide. The resuiting product isolated and purified in the manner of r ta Example 1D to yield l6/fi-n-butylpregnane-3a,17a-diol-1l, ZO-dione.

(E) 165 n butylpregnane 3a,l7oc,21 trial 11,20-

dione 21. acetate.-The 16,8-n-butylpregnane of Example 19D is reacted first with bromine in chloroform, then with sodium acetate and dimethyl-formam-ide and the product isolated and purified in the manner described in Example 1E,-to give 16,8-n-butylpregnane-3ot,170:,2l-triol-l1,20-dione ZI-acetate.

(F) 16;? n -butylpregnane 17a,21 dial 3,I1,20- trio/1e 21-acetate.-In the manner described in Example 1F, l6l3-n butylpregnane-3 t,l70L,2il'.l'iOl-l 1,20-dione 21- acetate is brominated and the resulting product isolated and purified to give 16,8-n-butylpregnane-17u,2l-diol-3,l1,

- 20-trione 2l-acetate.

(G) 4 bromo 16f n butylpregnane 170:,21 diol- 3,11,20 trione 21 acetate.1 6fi-n-butylpregnane-17a, 21-diol-3,11,20-trione 21-acetate is brominated in the man'- ner of Example 1G, to obtain 4-bromo-16B-n-butylpregnane-17u,2l-diol-3,1l,20-trione ZI-acetate.

- of aqueous alcoholic potassium bicanbonate in the manner described in Example 2 to yield IGb-n-butylcortisone,

Am? 238 m (615,000)

. Example 21 lfifi-METHYLPREDNISONE 21-ACETATE (A) 2,4-dibr0m0-16fl-methylpregnane 1700,21 -'di0l- 3,11,20-tri0ne 21-acetate.A' solution of 167 mg. 16;?- methylpregnane- 17a,2l-diol-3,11,20-trione 21-acetate, the compound of Example 1F, in 3 ml. of dioxane is dibrominated in positions 2 and 4 by the rapid addition of 130 mg. of bromine in 1 ml. of dioxane at room temperature. The solution is poured into water and the precipitated solid is filtered, yielding 180 mg. of 2,4-dibromo-16fimethylpregnane-17u,21-diol-3,11,20 trione 2l-acetate of suflicient purity to be used in the conversion described in the following procedure.

(13) .16 8-m ethylprednison'e 21 -acemte.-The 2,4-dibromide (180 mg.) as prepared in above Example 21A is dehydrobrominated by refluxing for 2 hours with 4 m1. of dimethylformamide containing 30 mg. of calcium carbonate. The mixture is poured into dilute hydrochloric acid and extracted with methylene chloride. extract is evaporated toa residue (150 mg), which is chromatographed on Florisil. The fractions obtained by elutionwith 20% ether-hexane are crystallized from ace- The organic tone-hexane to give .40 mg. of 16/3-methylprednisone 21- aceta-te,.M.P. 210-216". I One further crystallization gave x2433? 237 11,. 613.500.) Example 22 16B-METHYLPREDNISONE 16,8-methylprednisone 21 acetate (0.5 g.), the com pound of Example 21, when hydrolyzed by. means of 2 aqueous alcoholic potassium bicarbonate in the manner described in Example 2 yields lfifl-methylprednis'one, An alternative method of the preparation of the compound of this example is as follows: I

Bacillus sphaericus var. fusiformis A.T.C.C. 7055) is incubated on a nutrient agar'(compose'd'of Bacto-beef extract, 3 gQ; Bacto pe'ptone, 5 g.; sodium chloride, 8 'g.;

agar, 15g; tap Water, 1 liter) for 24 hours at 28 C. a

-To ml. of a sterile nutrient broth (composedof 7 20 Bacto-Beef extract, 3 g.; Bacto-peptone, 5 g.; per liter of tap water) in a 300 ml. flask is added one loopful 'of the incubated culture and the broth mixture is further incubated for 24 hours at 28 Cjon a shaking machine. The broth culture so obtained isemployed as an inoculum (1% Into each of ten flasks containing 100 ml. of sterile nutrient broth is added 1 ml. of the inoculum. The flasks are agitated on a rotary shaker for 8 hours at 28 C. at 240 strokes per minute. After this growth period, a solution of 25 mg. of 16,8-methylcortisone, the compound of Example 2, in 0.5 ml. of methanol is aseptically added to each flask which in turn is re-shaken and incubated for an additional 24 hours. The final pH is 7.8.

The contents of the flasks are then combined and extracted three times with Zliters of chloroform per extraction. The combined chloroform extracts are evaporated to dryness yielding 310 mg. of crude product. The crude" steroid is purified by chromatography on a chr0- matographic system described by G. M. Shull, Abstracts of Papers of the 126th Meeting of the American Chem- -ical Society, December 12-17, 1954, page 9 a, paper No.

24. Chromatographic evaluation shows a quantitative conversion of the starting material to the diene when an authentic sample of the 16fl-methylprednisone is used as a control.

Alternatively, the crude product is recrystallized from acetone affording 225 mg. of l6fl-methylprednisone.

Example 23 16/3-METHYLPREDNISOLONE. 16fi-methylhydrocortisone, the compound of Example 3, is fermented by meansof Bacillus sphaericus var. fusiformis (A.T.C.C. 7055) in the manner described in the alternate procedure of Example 22 to produce 16,8-methylprednisolone.

' r Example 24 16,6-METHYLPREDNISOLONE 2l-ACETATE l6p-methylprednisolone of Example 23, is treated with acetic acid and pyridine in the manner described in Example 4 to yield l6fi-methylprednisolone 21acetate.

Example 25 IGwMEIHYLPREDNISONE ZI-ACETATE concentrated hydrochloric acid. The mixture is extracted with methylene chloride, the organic extracts are washed with dilute sodium bicarbonate solution and water, then evaporated to a residue which is chromatographed on Florisil to give 15 mg. of l6ot-methylprednisone 21-acetate, A max. 238 m, (5 14,000

' Example 26 V .IGa-WIETHYLPREDNISONE In the manner described in Example 6, 03 g. of 16::-

aqueous alcoholic potassium bicarbonate to produce methylprednisone.

Example 27" I v V isa-l rur'nrwnnomsonn ZI-PROPIONATE' 16a-rnethylprednisone, the compound ofExample 26,

is reacted with propionic anhydride in pyridine in-the manner of Example 4.to. yield l6ct-methylprednis'one 21- I propionate. I

methylprednisone 21-acetate ishydrolyzed by means of- 2 Example 28 IGa-METHYLPREDNISOLONE Bacillus sphaericus var. fusiformis (A.T.C.C. 7055) is incubated on a nutrient agar (composed of Bacto-beef extract, 3 g., Bacto-peptone, 5 g., sodium chloride, 8 g., agar, 15 g., tap water, 1 liter) for 24 hours at 28 C.

To 100 ml. of sterile nutrient broth (composed of Bacto-beef extract, 3 g., Bacto-peptone, 5 g., per liter of tap water) in 300 mlflask is added one loopful of the incubated culture and the broth mixture is further incubated for 24 hours at 28 C. on a shaking machine. The broth culture so obtained is employed as an inoculum (1% Into each of ten flasks containing 100ml. of sterile nutrient broth is added 1 m1. of the inoculum. The flasks are agitated on a rotary shaker for 8 hours at 28 C. at 240 strokes per minute. After this growth period, a solution of 25 mg. of l6a-methylhydrocortisone, the compound of Example 7, in 0.5 ml. of methanol is aseptically added to each flask which, in turn, is reshaken and incubated'for an additional 24 hours. u The final pH is 7.8.

The contents of the flasks are combined and extracted three times with 2 liters of chloroform per extraction. The combined chloroformextracts are evaporated to dryness yielding 310 mg. of a-residue 'whichis crystallized from acetone-hexane to give 16a-methy1prednisolone.

Example 29 1Ga-M THYLPREDNISOLONE ZI-ACETATE A solution of 100 mg. of l6a-methylprednisol one in 2 ml. of pyridine containing 100 mg. of acetic anhydride is allowed to stand for 1 hour. The mixture is then poured into ice-HCl and a precipitate results which is re moved by filtration. Crystallization of the precipitate from aqueous methanol gives l6a-methylprednisolone 21- acetate.

Example 30 9a-BROMO-1Gfi-METHYLPREDNISOLONE ZI-ACETATE (A) 16,8 methyl 1,4,9(11)-pregnatriene-1 70;,21-di0l- 3,20-dine 21-acetate.A solution of 0.5 g. of 16B-methylprednisolone 2l-acetate, the compound of Example 24, in 3 m1. of pyridine is reacted with 0.3 ml. of methanesulfonyl chloride in 4 ml. of pyridine. The solution is allowed to stand for 6 hours, then poured into ice-hydrochloric acid. A 'solid precipitate which is removed by filtration and crystallized from acetone-hexane to give 16,8- methyl-1,4,9(11)-pregnatriene-l7u,21-diol-3,20-dione 2 lacetate,

MES 238 m (614,750)

(13) 90c bromo 16/3-mezhylprednis0lone 21-acetate. A suspension of 0.5 g. of the 16,8-methylpregnatriene of above Example 30A in 50 ml. of purified dioxane in a ml. of water containing 0.2 g. of N-bromoacetamide and 2 ml. of 1.5 N perchloric acid is gently agitated for a period of two hours, during which time the mixture becomes homogeneous. A solution of 0.5 g. of sodium sulfite in 5 ml. of water is added, and the reaction mixture is'extracted with methylene chloride. The organic extracts are washed with water, dried over magnesium sulfate, filtered and evaporated The resulting solid is crystallized from acetone to yield 0.3 g. of 9u-brotnod6fl-methylpregnisolone 2l-acetate,

I Example .31' Qa-FLUCRO-lGB-METHYLEREDNISOLONE 21-ACETATE (A) 9 8,1] 5 oxid0-16/3-methyl-1,4-pregnadiene-1 70421- di0l-3,20-di0ne 21-acetate.0ne gram of 9a-bromo-l6fimethylprednisolone 2l-acetate, the compound of Example 30, is added to 65 inl. of methanol and 1.0 g. of potassium acetate and the mixture is refluxed for 2 hours, then concentrated to a residue. Water is added to the residue and from methanol-water to yield 0.6 g. of 9fi,11 3-oxido-16fl- I methyl-1,4-pregnadiene-17a,21-diol-3,20-dione 2l-acetate,

(B) 902 fluoro-l6 8-methylprednisolone 21-acetate.A solution of 0.4 g. of the 93,1lp-oxido-lA-pregnadiene of Example 31A in 20 ml. of alcohol-tree chloroform is saturated with anhydrous hydrogen fluoride at 0 C. The mixture is allowed to stand 4 hours at 0 C., then concentrated to a residue under reduced pressure. Crystallization of the residue from acetone-hexane gives 0.2 g. 90:- fluoro-l6fi-methylprednisolone 2l-acetate,

my 238 mp (615,100) Example 32 9a-FLUORO-l6B METHYLPREDNISOLQNE Qa-BROMO-l Gfi-METHYLPREDNISOLONE 9a-bromo-16 8-methylprednisolone 2l-acetate, the com-- pound of Example 30,.is-. converted to 9a-bromo-l6fimethylpredn-isolone in the manner of Example 13 using hydrochloric acid in methanol-chloroform-water.

l 7 Example. 34 9wCHLORO-l6/i-l\EUJHYLPREDNISOLONE 21-ACETA1E A solution of 0.3 g. of 95,1lfi-oxido-lSli-methyl-IA- pregnadiene-l7a,2l-diol-3,20-dione 2l-acetate, the compound of Example 31A, in 40 ml. of alcohol-free chloroform is saturated at 0 C. with anhydrous hydrogen chloride, and the mixture allowed to stand at 0 C. for 6 hours. The mixture is concentrated under reduced pressure to a residue which is crystallized from acetone-Water to give .then idluted with water and extracted with methylene chloride. The organic extracts are washed with Water, dried over magnesium sulfate, filtered and evaporated in a residue which is crystallized from methanol to give 0.1 g. of 9a-fluoro16,8-methylprednisone ZI-acetate.

' Example 37 9a-FLUORO-16fi-METHYLPREDNISONE In the manner described in Example 13, 9a-fluoro-l618- methylprednisone 21-acetate is hydrolyzed to 9a-fluorol6fi-methylprednisone by means of HCl in methanolchloroform-water.

t i Example 38 9a-CHLORO-1Gfi-METHYLPREDNISONE (A) 906- chloro-l6,8-methylpr ednisone 21-acetate.9achloro-16 3-methy1prednisolone ZI-acet-ate, the compound of Example 34, is reacted with chromium rtioxide in aqueous acetic acid in the manner of Example 36 to give 9. ohloro-l6fi-methylprednisone 2l-acetate.

9u-chl0r0-16fl methyIpredrzisone-In the manner Y as described in Example 13, 38A; is reacted with hydrochloric acid in methanol-chloroform-water to give 9oa-chloro lfifmethylprednisone.

Example 39 I Qa-BROMO-16fl-METHYLPREDNISONE (A) 990 bromo 16,8-methylprednisne 21 acetate-'- 9ix-bromo 16p methylprednisolone 21-acetate, the compound of Example 30, is reacted with chromium trioxide in aqueous acetic acid in the manner of Example 36 to 7 give 9 tx-bromo-l6,8-methylprednisone 21-acetate.

(B) 9a'-br0m0-1 6,8-methylprednis0ne.-In the manner of Example 13, the 21-acetate ester of Example 39A is reacted with hydrochloric acid in methanol-chloroformwater to give 9 -bromo-16/3-methylprednisone.

. Example 40 9a-BBOMO-lGa METHYLPREDNISOLONE (A) 16oz methyl 1,4,9(11')-pregnatriene-17a,2l-di0l- 3,20-a'ion'e 2 1-acetate.-A solution of 0.3 g. of 16oamethy1- prednisolone 21-acetate, the compound of Example29, in

them-acetate ester of Example are-acre 2 ml. of pyridine is reacted with 0.1 ml. of methanesulfonyl chloride in 2 ml. of pyridine. The solution is allowed to stand for five hours,. then poured into icehydrochloric acid. A solid precipitates which is filtered and crystallized from acetone to give 16a -methyl- 1,4,9 1 1 pregnatriene-17a,21-diol-3,20-dione Zl-acetate,

W 238 my, 515,200)

(B) :9a-br0m0-1dot-methylpredizisolone 21-acetate.-A suspehsion of 0.3 g. of the 16a-methyl-1',4,9(11) pregnatriene' of Example 40A in ml; of purified dioxane, 3 ml. of water containing 0.1 g. of N-bromoacetamide and 1 ml. of 1.5 N perchloric acid is gently. agitated for a period of two hours, during which time the mixture becomes homogeneous. A solution of 0.3 g. of sodium sulfite in S'ml. of water is added, then the solution extracted with methylene chloride. The organic extracts arewashed with water, dried over magnesium sulfate,

filtered and evaporated. The resulting solid residue is crystallized from methanol to yield 0.2 g. of'9tx-bromo- 16u-methylprednisolone ZI-acetate,

P 243 m (e14,800)

(C) 9oz-bromo-loat-mthylprednis0l0ne.ln the mannor of Example 13, 9ot-bromo-16a-methylprednisolone 21f;

acetate is reacted with hydrochloric acid in chloroformmethanoi-water to give 9w-bromo-16u-methylprednisolone.

'Example 41 1 ,9a-ELUOR0 1(Sq-METHYLPREDNISOLONE (A) 95,11fi-oxid0-16u-methyl 1,4- pregnadiene-Uoa, 21-di0l-3,20-di0ne ZI-aCetateF-A mixture of 0.5 g. of 9a bromo-l6og-methylprednisolone 2l-acetate, the compound of Example B, 30 ml. of methanol and 0.5 g. of potassium acetate is refiuxedfor 2 hours, then concentrated to a residue. Water is added to the residue and the resulting solid isfiltered and crystallized from methanol to give 0.3 g; of 95,1lfl-oxido-16oa-inethy11,4-pregnadiene- 17a,21-diol-3,20-dione 21-acetate, 1 v

V 7 w 243m 14,050)' (B) 9u-flu0r0-16a-methylprednis0l0ne 21 acetate.

max.

A solution of 0.2 g. of the 96,1lfi-oxido-lM-methylpreg nadiene of above Example 41A in- 10' ml. of alcohol-free C.,- then concentrated to a'residue under reduced pressure.

I Crystallization of the, residue from acetone-hexane gives a 0.15 g. V of 9a-fiuoro loa-methylprednisolone ZI-acetate,

' hen max.

v 238 m (615,200) C) 96: -fluoro-ldoc-mllzylprednisolone. According f to the procedure of Example Busing hydrochloric acid in chloroform-mothanol-water, 9u-fiuoro 16st methylprednisolone.

. as Y prednisolone 21-acetate is converted to 9ot-fluoro-l6a methylprednisolone. I

Example 42 9aCHLOBO-lGa-METHYLPREDNISOLONE '(A) Qa-flzloro-l6a-methylprednisol0ne 21 acetate.

A solution of 0.2 g. of the 95,11,8-oxido-pregnadiene of Example 41A in 30 ml. of alcohol-free chloroform is saturated at 0 C. with anhydrous hydrogen chloride, and the mixture allowed to stand at this temperature for 6 hours. The mixture is concentrated under reduced pressure to. .a residue which is crystallized from acetonehexane to give 0.15 g. of 9a-chloro-16a-methylprednisclone '2 1 acetate.

Me OH MM.

I To a solution of 0.2 g. of 9a -fluoro-l6a-methylpredniso- ,lone ZI-ace'tate, the compound of Example 413, 10 ml.

of acetic acid there is addeddropwise a solution of 40 mg.

i of chromium trioxide. in 1 mLof Water and 2 ml. of acetic r chloroform is saturated with anhydrous hydrogen fluoride 1 at 0 C. The mixture is allowed to stand 5 hours at 0 acid. The resulting mixture-is allowed to standdhours, then diluted with water and extracted with methylene chloride. 'The organic extracts are washed with Water, dried over'magnesium sulfate, filtered and evaporated to a residue which is crystallized from methanol to give 0.1 g. of 9ot-fluoro-loot-methylprednisone 21-acetate,

A155? 237 mp. (614,000) (B) Qa-fluoro -Zdoc-methylprednisoneE-ln the manner of Example 13, the 21- acetate ester of Example 43 is hydrolyzed to C-rflllOI'O-16Ot7II1CthylPfdnlSOnQ Example 44 9wCHLORO-l6a-METHYLPREDNISONE In the manner described in Example 43, 906-0111010-1606- methylprednisolone 21-acetate, the compound of Example 42A, is reacted with chromium trioxide to yield 9achloro-16 x methylprednisone 2 1-.acetate.

In the manner described in Example 13, the 21-acetate ester of the compound of this example is reacted with hydrochloric acid in chloroform-mothanol-water to yield 90:-

, chloro-16u-met-hylprednisone.

Example 45 9aIBROMO-lGa-METHYLPREDNISONE In the manner described in Example 43, QOL-bI'OmO-IGOL- 1 methylprednisolone ZI-acetate, the compound of Example 40B, is reacted with chromium trioxide to yield 9a-bromo- 16ot-methylprednisone ZI-acetate. p T

In turn, the 21-acetate of the compound of this example is converted to the free alcohol, 9ot-bromo-16ot-methylprednisone, by means oat-hydrochloric acid in chloroformmethanol-water according to the procedure of Example 13.

Example 46 9a-CH LORO-16a-METHYLHYDROCORTISONE; In the manner described in Example 42, 95,11f3-oxido- 16a-methyl-4spregnene-17cx,21-di0l 21-acetate, the compound of Example 18A, is reacted with hydrogen chloride in chloroform to give 9ot-chlo-ro-16a-methylhydro- .cortisone 21-acetate,

The 21-acetate of the compound of this example is hydrolyzed by means of hydrochloric acid in chloroformmethanol-water according to the procedure 'of Example 13, to obtain 9a-ch1oro-16u-methylhydrocortisone.

Example 47 9a-CHLORO-16a-METHYLCORTISONE In the manner described in Example 36, 9a-chloro- 16a-methylhydrocortisone 21-acetate is reacted with chromium trioxide in water and acetic acid" to yield 9achloro-lfia-methylcortisone 21 acetate,

W 239 my 15,100

The 21-acetate of the compound of this example is hydrolyzed by means of hydrochloric acidin chloroformmethanol-water according to the procedure of Example 13 to give 9a-chloro-16a-methylcortisone.

Example 48 Sa-FLUORO-lGa-IWETHYLCORTISONE' M MA...

237 my, (e15,000)

In the manner of Example 13, the 21-acetate of the compound of this example is hydrolyzed with hydrochloric acid in chloroform-methanol-water to yield 90:-

fluoro-16a-methylc0rtisone.

Example 49 9a-BROMO-16a-ME-THYLCORTISONE In the manner described in Example 36, 9a-bromo-16amethylhydrocortisone ZI-acetate, the compound of Example 17B, is reacted with chromium tn'oxide in aqueous acetic acid to give 9a-bromo-lGa-methylcortisone 21-acetate,

The 21-acetate of the compound of this example is hydrolyzed by means of hydrochloric acid in methanolwater-chloroform in the manner described in Example 13 to give 9a-bromo-l6a-methylcortisone.

Example 50 lfia-ETHYLCORTISONE 21-ACETATE By performing the sequence of reactions described in Example 9 (A-F), l6-pregnene-3a-ol-l1,20-di0ne is first reacted with ethyl iodide and magnesium to yield the requisite intermediate, 16a-ethylpregnane 3oz ol 11,20- dione which, in turn, is ultimately converted to 16a-ethylcortisone 21-acetate which compound is isolated and purified in the manner described in Example 9F.

16a-ethylhydrocortisone is subjected to the action of Bacillus sphaericus var. fusiformis and the resulting prod- 26 net isolated and purified in the manner described in Example 22 to give 16a-ethylprednisolone.

The ZI-acetate of 16u-ethylprednisolone is prepared from the free alcohol and acetic anhydride and pyridine in the manner described in Example 29.

Example 55 9a-BROMO-lGa-ETHYLPREDNISOLONE 16a-ethylprednisolone 21-acetate is converted to bromo-l6a-ethylprednisolone 21-acetate and thence to the corresponding 21-alcohol by going through the sequence of reactions described in Example 4D.

Example 56 Qa-FLUORO-IGa-ETHYLPREDN IS OLONE The 9a-bromo-16a-ethylprednisolone ZI-acetate of Example 55 is converted to 9,3,11fi-oxido-16a-ethylprednisolone and thence to 9a-fluoro-16a-ethylprednisolone 21- acetate and the corresponding 21-alcohol in the manner described in Example 41.

Example 57 9a.-FLUORO-IGa-ETHYIXPREDNISONE 21-ACETATE In the manner described in Example 36, 9a-fluoro-16aethylprednisolone 21-acetate is reacted with chromium trioxide in aqueous acetic acid to give 9a-fluoro-l6aethylprednisone 21-acetate.

What is claimed is:

1. The pregnene having a keto group at the 3- and 20- positions, hydroxy at the 17a-position, a member of the group consisting of hydroxy and pharmaceutically acceptable esters thereof at the 21-position, a member of the group consisting of hydrogen and halogen at the 9aposition and characterized by a member of the group consisting of keto and S-hydroxy at the ll-position, a lower alkyl group at the 16/3-position and the presence of a double bond between positions selected from the group consisting of (4,5) and (1,2;4,5).

2. A compound of the formula:

zOR

wherein X is selected from the group consisting of hydrogen and halo; W is selected from the group consisting of keto and fl-hydroxy; and R is selected from the group consisting of hydrogen and lower alkanoyl.

3. A compound of the formula:

wherein X is selected from the group consisting of hydrogen and halo; W is selected from the group consisting of keto and B-hydroxy; and R is selected from the group consisting of hydrogen and lower alkanoyl.

. 16,6-lower alkyl cortisone.

. 16 3-lower alkyl hydrocortisone.

. 16,6-lower alkyl prednisone.

. 16/3-lower alkyl prednisolone.

. 9a-halogeno-16p-lower alkyl hydrocortisone.

27 9. 9a-ha10gen0-16p-lower alkyl prednisolone. 10. A 21-pharmaceutically acceptable ester of l6 8-lower alkyl cortisone.

11. A21-pharmaceutically acceptable ester of 16,8-1ower alkyl hydrocortisone.

12. A 21-pharrnaceutical1y acceptable ester of 16fl-lower alkyl prednisone.

13. A 21-pharmaceutically acceptable ester of 16B-1ower alkyl prednisolone. V

14. A 21-pharmaceutic'ally acceptable ester 'of 9a-halogen0-16B-l0wer alkyl hydrocortisone.

15. A 21-pharmac'eutically acceptable ester of 9zx-halogeno-16fl-l0wer alkyl prednisolone.

16. 16,8-methylprednisone.

17. 16/3-methylprednis0lone;

18. 9a-fluoro-16,8-methylprednisolone.

19. A 21 pharmaceutically acceptable ester of 16;?- methylprednisone.

. 20. A 21 pharmaceutically acceptable ester of 16 6- methylprednisolone.

21. A 21 pharmaceutically acceptable ester of 90afluoro-l6,8-methy1prednisolone.

22. 9a-fluoro 16fl-methylprednisolone 21-acetate.

23. The dihydrogen phosphate esters of the compounds of claim 1. 1

No references cited.

LEWIS GOTTS, Primary Examiner.

Notice of Adverse Decision in Interference In Interference No. 96,052 involving Patent No. 3,164,618, R. Rausser and E. P. Oliveto, ALKYLATED STEROIDS final udgment adverse to the patentees was rendered Apr. 4, 1969, as to claims 18 and 22.

[Oficz'al Gazette J1me 3, 1.969.]

Claims (1)

1. THE PREGNENE HAVING A KETO GROUP AT THE 3- AND 20POSITIONS, HYDROXY AT THE 17A-POSITION, A MEMBER OF THE GROUP CONSISTING OF HYDROXY AND PHARMACEUTICALLY ACCEPTABLE ESTERS THEREOF AT THE 21-POSITION, A MEMBER OF THE GROUP CONSISTING OF HYDROGEN AND HALOGEN AT THE 9APOSITION AND CHARACTERIZED BY A MEMBER OF THE GROUP CONSISTING OF KETO AND B-HYDROXY AT THE 11-POSITION, A LOWER ALKYL GROUP AT THE 16B-POSITION AND THE PRESENCE OF A DOUBLE BOND BETWEEN POSITIONS SELECTED FROM THE GROUP CONSISTING OF (4,5) AND (1,2;4,5).